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1.
Species of the genus Syphacia are considered to have generally co-evolved with their rodent hosts. This study determined partial sequences of the CO1 gene from several species in the genus Syphacia and discuss the relationships between pinworms and their hosts. Syphacia montana, which parasitizes Microtinae, was closely related to S. frederici and S. obvelata, which parasitize Murinae. Although both S. obvelata and S. ohtaorum parasitize rodents in the genus Mus, these two species were not found to be closely related to each other. Syphacia frederici, S. emileromani and S. agraria are all pinworms of the Apodemus species, but genetic affiliation between these three species was not indicated. These facts suggest that the co-evolutionary relationship between species of the genus Syphacia and their host rodents may not so strict and host switching has probably occurred during the course of evolution.  相似文献   

2.
郝桂英  何学谦 《中国畜牧兽医》2015,42(12):3167-3172
应用保守引物BD1和BD2对7个鸡蛔虫凉山州分离株的核糖体DNA内转录间隔区(ITS)及5.8S rDNA序列进行PCR扩增和序列测定,并用ITS-1、ITS-2序列重构鸡蛔虫与其他蛔虫的系统发育关系。测序结果显示所获得的鸡蛔虫ITS及5.8S rDNA序列大小为974~989 bp,同源性为98.9%~100.0%。其中ITS-1、5.8S rDNA和ITS-2片段大小分别为473~481、157和337~359 bp,同源性分别为98.5%~100.0%、100.0% 和98.5%~100.0%。系统发育树显示所有鸡蛔虫分离株聚在同一分支,能与其他蛔虫相区别。研究结果表明,鸡蛔虫的ITS-1、ITS-2序列种内变异小,但种间差异大,故可作为分子标记用于鸡蛔虫的虫种鉴定,为鸡蛔虫的分子分类、分子流行病学调查和种群遗传的进一步研究奠定基础。  相似文献   

3.
安氏隐孢子虫ITS-1序列的PCR扩增、克隆及分析   总被引:1,自引:0,他引:1  
通过对国内三株安氏隐孢子虫(Cryptosporidium andersoni)即GD株、HN株和AH株的rDNA的内转录间隔区Ⅰ(ITS_1)序列进行PCR扩增、克隆、测序和序列分析,旨在确定ITS_1是否可作为C.andersoni分子分类的遗传标记。结果表明:GD株、HN株和AH株的ITS_1序列基本一致,仅AH株有三个碱基的差异;但与GenBank注册的C.muris和C.parvum存在种间差异,而且差异显著。说明ITS_1可作为C.andersoni种的遗传标记,从而为隐孢子虫属的种间鉴定以及进一步的分子流行病学调查和分子诊断学研究奠定了基础。  相似文献   

4.
根据OenBank上已发表的环孢子虫(Cyclospora)rDNA序列设计并合成1对引物,利用PCR技术时首次在牛体内发现的形态学特征与人环孢子虫(Cyclospora cayetanensis)极为相似的牛源环孢子虫的ITS-1+序列进行了扩增。将PCR扩增出的片段纯化后,克隆至pGEM—TEasy载体后进行序列测定,并利用NCBI在线BLAST程序对测序结果进行了同源性比较和序列分析。分析结果显示,扩增的ITS-1+大小为865bp的片段,包含18S部分序列(371bp)、ITS-1全序列(385bp)和5.8S部分序列(109bp)。序列同源性分析表明,该牛源环抱子虫为艾美尔科原虫,但不同于目前已知的各种艾美尔科原虫,可能是一种新发现的原虫。  相似文献   

5.
The diagnosis of tapeworm infections in horses relies on copro-diagnostic methods, which are time-consuming and of limited sensitivity for determination of the exact prevalence. The development of serological tests has slightly improved the detection of tapeworm infections, but more sensitive methods are still required. A polymerase chain reaction (PCR)-based approach may constitute a valuable tool to improve tapeworm diagnosis. Nuclear ribosomal DNA (rDNA) is a useful target for species and/or strain markers. Partial 18S, the internal transcribed spacer 1 (ITS-1), the 5.8S, the internal transcribed spacer 2 (ITS-2), and partial 28S rDNA of the equine tapeworms Anoplocephala perfoliata and Anoplocephaloides mamillana were amplified and sequenced. The lengths and GC contents of the regions sequenced were 2087-2091bp and 49.35-49.69% for A. perfoliata, and 2110-2119bp and 49.15-49.32% for A. mamillana, respectively. Sequence alignment and comparison of both taxa showed 79.3-80.2% identity. The lowest identities were found in the ITS regions with 39.9-43.5% for the ITS-1 and 59.5-61.2% for the ITS-2. No matches of the ITS-2 of A. perfoliata and A. mamillana were found with other species by BLAST search. For this reason, ITS-2 sequences seemed appropriate as accurate species markers and A. perfoliata ITS-2 primers were developed. The ITS-2 PCR enabled the detection of genomic DNA as low as 0.5 pgs. First efforts on the practical application of the PCR-based approach were made. A 6-mg fragment of a tapeworm proglottid was detected in 0.5 and 1g of faeces.  相似文献   

6.
对从试验小鼠体内分离的蛲虫样品PMZ1-5的线粒体细胞色素c氧化酶第I亚基(cox1)的部分基因(pcox1)进行PCR扩增、测序及序列分析,并与网上相关序列进行比对分析,研究其线粒体pcox1基因遗传变异情况。结果表明,获得pcox1有效序列373 bp,经序列比对分析,发现5个蛲虫样品之间的变异很小,只有1个碱基的差异,序列相似性为99.73%~100%,BLAST分析结果表明为隐藏管状线虫,与同属的Syphacia montana序列差异性为5.24%~5.68%。结果表明,隐藏管状线虫的pcox1序列种内变异很小,种间差异明显,本结果为进一步研究蛲虫的群体遗传学奠定了基础。  相似文献   

7.
本试验首次研究了小鼠隐藏管状线虫及四翼无刺线虫在线粒体烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶亚单位5基因(nad5)部分序列(pnad5)的种间及种内变异。通过对甘肃兰州地区实验小鼠体内分离的蛲虫样品(LY1-20、LS1-22) nad5基因pnad5进行PCR扩增、克隆及测序分析,研究其遗传变异情况。结果表明,获得隐藏管状线虫pnad5有效序列753 bp,通过序列比对发现来自LY和LS 2家实验动物中心的22个隐藏管状线虫样品之间只存在3个碱基的差异,序列相似性99.60%。获得四翼无刺线虫pnad5有效序列865 bp,2大动物中心20个四翼无刺线虫样品之间没有碱基的差异,序列相似性100%。用软件ClustalX 1.83截取2种鼠蛲虫nad5基因长度共同的序列,通过比对发现2种鼠蛲虫种间差异为53.64%,种间差异显著。研究结果表明,线粒体nad5基因在鼠蛲虫种内相对保守,但种间有着较高的突变率,可作为鼠蛲虫种间鉴定、诊断的遗传标记。  相似文献   

8.
In the present study, samples representing Bunostomum trigonocephalum and Bunostomum phlebotomum from sheep and cattle in Heilongjiang Province, China, were characterized and grouped genetically by the first (ITS-1) and second (ITS-2) internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA). The rDNA region including the ITS-1, 5.8S, ITS-2, and flanking 18S and 28S rDNA sequences was amplified by polymerase chain reaction (PCR), then sequenced and compared with that of other members of the hookworms available in GenBank?, and phylogenetic relationships between them were reconstructed using the Maximum-Parsimony method. The ITS-1, 5.8S, and ITS-2 sequences of the sheep hookworm were 381, 153, and 231 bp in length, respectively, and the corresponding sequences of the cattle hookworm were 392, 153, and 240 bp in length. The identity of ITS sequences of B. trigonocephalum and B. phlebotomum from sheep and cattle was 87.4%. A PCR-linked restriction fragment length polymorphism (PCR-RFLP) assay using restriction endonuclease Nde I was established for the unequivocal differentiation of the two hookworm species. Phylogenetic analyses based on the ITS sequences revealed that B. trigonocephalum and B. phlebotomum were closely related, but they represent two different species.  相似文献   

9.
The present study examined the relationship among individual Sarcoptes scabiei mites from 13 wild mammalian populations belonging to nine species in four European countries using the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA (rDNA) as genetic marker. The ITS-2 plus primer flanking 5.8S and 28S rDNA (ITS-2+) was amplified from individual mites by polymerase chain reaction (PCR) and the amplicons were sequenced directly. A total of 148 ITS-2+ sequences of 404 bp in length were obtained and 67 variable sites were identified (16.59%). UPGMA analyses did not show any geographical or host-specific clustering, and a similar outcome was obtained using population pairwise Fst statistics. These results demonstrated that ITS-2 rDNA does not appear to be suitable for examining genetic diversity among mite populations.  相似文献   

10.
In this study, the effect of doramectin, moxidectin and netobimin was investigated in naturally infected Syphacia muris in rats. The natural infection was determined by the use of cellophane tape method on the perianal region and by the technique of centrifugal flotation of feces. The infected rats were divided into three treated and one control group (N = 10). Doramectin and moxidectin at the dose of 0.2 mg/kg per day and netobimin at the dose of 7.5 mg/kg per day were given in the diet for 4 days. Cellophane tape preparations were performed in all groups on 4th and 7th day after the last treatment. The rats of treated groups were necropsied on 7th day after the last treatment together with that of control group. While doramectin and netobimin were highly effective against S. muris, moxidectin was not found to be effective for eradication of S. muris.  相似文献   

11.
Streptococcus dysgalactiae serogroup C, G and L strains were investigated by polymerase chain reaction (PCR) using oligonucleotide primers designed according to species-specific parts of the 16S-23S rDNA intergenic spacer region. The oligonucleotide primers with specificity for the 16S-23S rDNA intergenic spacer region allowed a correct identification of all S. dysgalactiae serogroups C, G and L strains investigated. No cross-reactivities could be observed with any of the control strains indicating the usefulness of PCR-technology to identify the serologically heterogeneous species S. dysgalactiae.  相似文献   

12.
从河北省兔场分别单卵囊分离孢子化大型艾美耳球虫卵囊、黄艾美耳球虫卵囊及肠艾美耳球虫卵囊,接种无球虫兔后获得纯种卵囊,CTAB法提取孢子化卵囊基因组DNA。利用艾美耳属球虫18SrDNA和5.8SrDNA保守引物,PCR扩增3种兔球虫ITS-1片段,产物纯化后测序。将3种球虫ITS-1测序结果与GenBank发布的兔球虫ITS-1序列进行比对和遗传距离比较,绘制系统发育树。结果表明,大型艾美耳球虫、黄艾美耳球虫及肠艾美耳球虫河北株分别扩增出424、455、434bp的ITS-1片段。大型艾美耳球虫、黄艾美耳球虫及肠艾关耳球虫河北株与GenBank中发布的同种兔球虫ITS-1序列相似性分别为97.4%、97.9%和96.9%。系统发育树显示兔球虫ITS-1序列形成1个单系群,该单系群根据寄生部位分为2个姊妹群。  相似文献   

13.
美洲狮和亚洲黑熊蛔虫ITS序列扩增及序列分析   总被引:1,自引:1,他引:0  
以采自广州动物园美洲狮和亚洲黑熊体内的蛔虫样品为研究对象,提取样品的总DNA,以保守引物对核糖体DNA内转录间隔区(ITS)序列进行PCR扩增、测序及网上比对。结果表明,美洲狮体内样品获得与GenBankTM公布的狮弓首蛔虫相似性很高的ITS-1、ITS-2和5.8S序列,亚洲黑熊体内样品获得与GenBankTM公布的转移拜林蛔虫相似性很高的ITS-2序列,并首次获得样品的ITS-1与5.8S序列。序列分析结果表明,美洲狮体内蛔虫样品为狮弓首蛔虫,亚洲黑熊体内蛔虫样品为转移拜林蛔虫。  相似文献   

14.
猪食道口线虫ITS-1和lTS-2 rDNA的PCR-SSCP分析   总被引:1,自引:0,他引:1  
以采自我国不同地区猪体的食道口线虫虫株为研究对象,PCR扩增出ITS-1和ITS-2序列片段,然后采用单链构象多态性(SSCP)方法分析PCR产物,对不同地区食道口线虫进行分子鉴定。所有样品经SSCP分析显示两种带型,第一种为有齿食道口线虫带型,另一种为未定种食道口线虫带型。代表性样品的测序结果表明,未定种食道口线虫带型的样品为四棘食道口线虫。本研究在国际上首次报道了中国猪四棘食道口线虫的ITS序列,并建立了区分有齿食道口线虫和四棘食道口线虫的PCR-SSCP方法,从而为食道口线虫的分子生物学的进一步研究奠定了基础。  相似文献   

15.
Isolates of the rumen fluke Calicophoron daubneyi (Digenea: Paramphistomidae) from various hosts and three locations in southern Italy were characterized genetically. The second internal transcribed spacer (ITS-2) of ribosomal DNA (rDNA) plus flanking 5.8S and 28S sequence (ITS-2+) was amplified from individual rumen flukes by PCR. PCR-linked restriction fragment length polymorphism (PCR-RFLP) analysis was performed using four different restriction endonucleases, and PCR products were sequenced. The PCR analyses from all the C. daubneyi specimens produced identical fragments, and the PCR-RFLP analyses did not show, with respect to any of the four restriction endonucleases, any differences between the C. daubneyi specimens. The sequence analyses of the ITS-2+ from each of the C. daubneyi specimens showed them all to be 428 bp, and composed of the entire ITS-2 sequence (282 bp) plus the two partial flanking conserved sequences, 5.8S (99 bp) and 28S (47 bp). No intra-specific variation was observed in the nucleotide composition of the ITS-2+ (homology=100%). There was, however, an observable inter-specific variation between the ITS-2+ of C. daubneyi and the ITS-2+ of both Calicophoron calicophorum (homology=97.2 %) and Calicophoronmicrobothrioides (homology=97.4 %), both previously deposited in the GenBank. The finding of the present study shows that, as has already demonstrated for other parasitic helminths, ITS-2 can serve as an effective genetic marker for the molecular identification of paramphistomes, and as a useful tool for developing molecular epidemiological techniques for the study of C. daubneyi transmission patterns and prevalence in definitive and intermediate hosts.  相似文献   

16.
本研究旨在阐明黄鳝胃瘤线虫湖南分离株的核糖体DNA(rDNA)内转录间隔区(ITS)及5.8 S rDNA序列的遗传变异情况,并用ITS序列重构胃瘤线虫与其它线虫的种群遗传关系.利用聚合酶链反应(PCR)扩增胃瘤线虫rDNA的ITS-1、5.8S及ITS-2片段,将PCR扩增出的片段纯化后克隆至pGEM-T Easy载体,重组质粒通过菌落PCR鉴定后,对阳性菌落进行序列测定并进行序列分析.结果显示所获得的胃瘤线虫ITS及5.8 S rDNA序列总长存在一定差异(922~927 bp),其中包含部分的18S、28 S及全部的ITS-1 (350~351 bp)、5.8S(102 bp)及ITS-2 (340~344 bp)序列.本研究系国内首次报道胃瘤线虫的ITS序列,其结果为黄鳝胃瘤线虫的分类鉴定以及进一步的分子流行病学调查和群体遗传研究奠定了基础.  相似文献   

17.
利用聚合酶链反应(PCR)扩增蛇蛔虫rDNA的ITS-1、5.8S及ITS-2片段,将PCR扩增出的片段纯化后克隆至pGEM-T Easy载体,重组质粒通过菌落PCR鉴定后,对阳性菌落进行序列测定并进行序列分析。结果显示,所获得的蛇蛔虫ITS及5.8SrDNA序列总长为846bp,包含部分的18S、28S及全部的ITS-1、5.8S及ITS-2序列。本研究系国际上首次报道蛇蛔虫的ITS序列,从而为蛇蛔虫的分类鉴定以及进一步的分子流行病学调查奠定了基础。  相似文献   

18.
以广州动物园小熊猫体内分离出的蛔虫为研究对象,运用PCR方法,以保守引物NC5和NC2扩增其核糖体DNA(rDNA)的内转录间隔区(ITS)和5.8S序列,并对扩增后的片段进行纯化、克隆至pGEM-Teasy载体、转化、测序和序列分析,以鉴定小熊猫蛔虫的种类。结果显示2条蛔虫样品的ITS及5.8S rDNA序列基本一致,总长为913 bp,样品间序列相似性为99.7%。将序列与GenBankTM公布的相关序列进行比较分析,结果显示2条蛔虫的ITS及5.8S序列与黑熊横走贝蛔虫(Baylisascaris transfuga注册号AB571304)相似性分别为98.1%、98.4%,与大熊猫西氏贝蛔虫(Baylisascaris schroederi注册号JN210912)相似性分别为96.9%、97.1%,与猪蛔虫(Ascaris suum注册号AB571302)相似性分别为89.9%、90.1%,与人蛔虫(Ascaris lumbricoides注册号AB571296)相似性分别为89.8%、90.1%,ITS-1序列与浣熊贝蛔虫(Baylisascaris procyonis注册号AB053230)相似性分别为92.0%、92.3%。研究结果表明小熊猫体内分离的蛔虫可能为贝蛔属蛔虫,从而为蛔虫的进一步分类、鉴定和遗传变异研究奠定了基础。  相似文献   

19.
基于核糖体DNA第一与第二转录间隔序列以及5.8S序列,以分离自广州动物园大熊猫体内的蛔虫为研究对象,用保守引物NC_5和NC_2对核糖体DNA(rDNA)的内转录间隔区ITS-1,ITS-2及5.8S序列进行PCR扩增,扩增后的片段纯化后克隆至pGEM-Teasy载体,重组质粒通过菌液PCR鉴定后,对阳性菌落进行序列测定及分析,鉴定大熊猫蛔虫的种类。结果显示,目的片段总长为910 bp,2个不同样品之间的ITS及5.8S序列没有差异,与GenBank~(TM)中的拜林蛔线虫(Baylisascaris transfuga)、猪蛔虫(Ascaris suum)和人蛔虫(Ascaris lumbricoides)的ITS序列相似性分别为96.6%、82.9%和82.7%。结果表明,此次分离的大熊猫蛔线虫可能为拜林蛔线虫。  相似文献   

20.
Although Spiculopteragia spiculoptera is primarily a parasite of cervids, it can also but less often contaminate domestic livestock. Little is known about its epidemiology and its pathogenicity in domestic ruminants and other unusual cervid species. Its taxonomic status remains unclear since the hypothesis of morphological polymorphism among males has been proposed. However, accurate taxonomy is fundamental in the identification and survey of potentially pathogenic species of parasites. The second internal transcribed spacer of rDNA (ITS-2) and the mitochondrial (mt) DNA-derived ND4 gene were used to study the polymorphism hypothesis for S. spiculoptera. ND4 evolves more quickly than ITS-2 and is considered to be more discriminant in the characterization of closely related species. DNA sequences of ITS-2 and ND4 were studied in 18 individual males of morphological type spiculoptera and in 3 of morphological type of mathevossiani from Red deer (Cervus elaphus), Roe deer (Capreolus capreolus) and Chamois (Rupicapra rupicapra). Intraindividual ITS-2 variations were detected within and between each morphotype of Spiculopteragia but these differences did not separate the two morphs mathevossiani and spiculoptera. Similarly, although ND4 showed a high level of nucleotide substitution, the morphotypes S. mathevossiani and S. spiculoptera were clustered together. Our genetic data support the dimorphic male hypothesis for the species S. spiculoptera.  相似文献   

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